Monitoring AKT activity and targeting in live tissue and disease contexts using a real-time Akt-FRET biosensor mouse

James R.W. Conway; Sean C. Warren; Young Kyung Lee; Andrew T. McCulloch; Astrid Magenau; Victoria Lee; Xanthe L. Metcalf; Janett Stoehr; Katharina Haigh; Lea Abdulkhalek; Cristian S. Guaman; Daniel A. Reed; Kendelle J. Murphy; Brooke A. Pereira; Pauline Mélénec; Cecilia Chambers; Sharissa L. Latham; Helen Lenthall; Elissa K. Deenick; Yuanqing Ma; Tri Phan; Elgene Lim; Anthony M. Joshua; Stacey Walters; Shane T. Grey; Yan Chuan Shi; Lei Zhang; Herbert Herzog; David R. Croucher; Andy Philp; Colinda L.G.J. Scheele; David Herrmann; Owen J. Sansom; Jennifer P. Morton; Antonella Papa; Jody J. Haigh; Max Nobis; Paul Timpson

doi:10.1126/sciadv.adf9063

Monitoring AKT activity and targeting in live tissue and disease contexts using a real-time Akt-FRET biosensor mouse

James R.W. Conway
, Sean C. Warren
, Young Kyung Lee
, Andrew T. McCulloch
, Astrid Magenau
, Victoria Lee
, Xanthe L. Metcalf
, Janett Stoehr
, Katharina Haigh
, Lea Abdulkhalek
, Cristian S. Guaman
, Daniel A. Reed
, Kendelle J. Murphy
, Brooke A. Pereira
, Pauline Mélénec
, Cecilia Chambers
, Sharissa L. Latham
, Helen Lenthall
, Elissa K. Deenick
, Yuanqing Ma

Tri Phan, Elgene Lim, Anthony M. Joshua, Stacey Walters, Shane T. Grey, Yan Chuan Shi, Lei Zhang, Herbert Herzog, David R. Croucher, Andy Philp, Colinda L.G.J. Scheele, David Herrmann, Owen J. Sansom, Jennifer P. Morton, Antonella Papa, Jody J. Haigh^*, Max Nobis^*, Paul Timpson^*

^*Korresponderande författare för detta arbete

Åbo biovetenskapscentrum

Forskningsoutput: Tidskriftsbidrag › Artikel › Vetenskaplig › Peer review

9 Citeringar (Scopus)

52 Nedladdningar (Pure)

Sammanfattning

Aberrant AKT activation occurs in a number of cancers, metabolic syndrome, and immune disorders, making it an important target for the treatment of many diseases. To monitor spatial and temporal AKT activity in a live setting, we generated an Akt-FRET biosensor mouse that allows longitudinal assessment of AKT activity using intravital imaging in conjunction with image stabilization and optical window technology. We demonstrate the sensitivity of the Akt-FRET biosensor mouse using various cancer models and verify its suitability to monitor response to drug targeting in spheroid and organotypic models. We also show that the dynamics of AKT activation can be monitored in real time in diverse tissues, including in individual islets of the pancreas, in the brown and white adipose tissue, and in the skeletal muscle. Thus, the Akt-FRET biosensor mouse provides an important tool to study AKT dynamics in live tissue contexts and has broad preclinical applications.

Originalspråk	Engelska
Artikelnummer	eadf9063
Antal sidor	15
Tidskrift	Science Advances
Volym	9
Nummer	17
DOI	https://doi.org/10.1126/sciadv.adf9063
Status	Publicerad - apr. 2023
MoE-publikationstyp	A1 Tidskriftsartikel-refererad

Finansiering

Acknowledgments:W ewouldliketothankthestaffatthefollowingfacilitiesattheGarvan InstituteofMedicalResearch:Austr alian BioResource(ABR),biologicaltestingfacility(BTF), tissueculturefacilityunderG.LehrbachandR.Lyons,theARCFINCIT ecenterunderco-leadershipofPhanandTimpson,andthehistologycorefacilityundertheleadershipof A. Zaratzian. W e would also like to thank C. Winchester for critical reading of the manuscript. SomeimageswereadaptedfromServierMedicalArt,licensedundertheCreativeCommons Attribution3.0UnportedLicense(https://creativecommons.org/licenses/by/3.0),andother schematicimageswerecreatedwithBioRender.com.Funding:Thisstudywassupportedby NationalHealthandMedicalResearchCouncil(NHMRC),Austr alian ResearchCouncil(ARC), CancerCouncilNSW ,CancerInstituteNSW ,CancerAustralia,NationalBreastCancerFoundation (NBCF), Tour de Cure, St Vincent’s Clinic Foundation, Tour de Cure and Sydney Catalyst (the Translational Cancer Research Centre of central Sydney and regional New South W ales), an AustralianCancerResearchFoundation(A CRF INCIT e) infrastructuregrant,andSuttonsfamily andLenAinsworthfoundationphilanthropy.J.J.H.wouldliketoacknowledgefundingbythe CIHRandCancerCareManitobaFoundation.P .T .isaNationalHealthandMedicalResearch Council(NHMRC)SeniorResearchFellow.M.N.andD.H.aresupportedbyCINSWEarlyCareer Research Fellowships and St Vincent’s Clinic Foundation grants and NHMRC Ideas grants. J.R.W .C. was supported by the European Union’s Horizon 2020 research and innovation programmeundertheMarieSklodowska-Curiegrantagreement(841973)andanAcademyof Finland postdoctoral research grant (338585). O.J.S. and J.P .M. were supported by Cancer ResearchUKcorefundingtotheBeatsonInstitute(A31287),toO.J.S.labora tory (A21139),and toJ.P .M. laboratory(A29996).B.A.P .andK.J.M.aresupportedbyCINSWEarlyCareerResearch Fellowships.B.A.P .issupportedbyaPanKind(PancreaticCancerFoundation)Australia,UNSW Medicine Cancer Theme, Perpetual Impact Philanthropy, and St Vincent’s Clinic Foundation grants.Y .-C.S. issupportedbyanNHMRCgrantandtheDiabetesAustraliaResearchProgram. T .P . is supported by a senior research fellowship through the National Health and Medical ResearchCouncil(NHMRC1155678).Authorcontributions:Investigation,validation,and formalanalysis:J.R.W .C., S.C.W ., Y .-K.L., A.T .M., C.C.,A.M.,V .L., X.L.M.,J.S.,K.H.,L.A.,C.S.G.,D.A.R., K.J.M.,B.A.P ., P .M., S.L.L.,H.L.,E.K.D.,Y .M., T .P ., E.L.,A.M.J.,S.W ., S.T .G., Y .-C.S., L.Z.,H.H.,A.Ph., D.R.C.,A.Pa.,M.N.,andP .T .Conceptualizationandfundingacquisition:J.R.W .C., S.C.W ., K.J.M., B.A.P ., C.L.G.J.S.,D.H.,O.J.S.,J.P .M., A.Pa.,J.J.H.,M.N.,andP .T .Writingandvisualization:M.N.and P .T . Competing interests: P .T . receives reagents from Kadmon Inc., INXMED (also consultant), CRUKAstraZenecaAlliancelabora tory ,RedXPharma,EqulibreBiopharmaceuticals,andAmplia Therapeutics.UnderalicensingagreementbetweenAmpliaTherapeuticsandGarvanInstitute ofMedicalResearch,K.J.M.,D.H.,andP .T .(consultant)areentitledtomilestonepayments. O.J.S. ’s laboratory receives funding from Cancer Research Horizons, Redex, AstraZeneca, and Novartis. J.P .M. ’s labora tory receives funding from AstraZeneca, UCB Pharma, and Redex. All otherauthorsdeclarethattheyhav enocompetinginterests.Dataandmaterialsavailability: Alldataneededtoevaluatetheconclusionsinthepaperarepresentinthepaperand/orthe SupplementaryMaterials.TheAkt-FRETmousecanbeprovidedbythecorrespondingauthors pending scientific reviewand a completed material transfer agreement. This study was supported by National Health and Medical Research Council (NHMRC), Australian Research Council (ARC), Cancer Council NSW, Cancer Institute NSW, Cancer Australia, National Breast Cancer Foundation (NBCF), Tour de Cure, St Vincent's Clinic Foundation, Tour de Cure and Sydney Catalyst (the Translational Cancer Research Centre of central Sydney and regional New South Wales), an Australian Cancer Research Foundation (ACRF INCITe) infrastructure grant, and Suttons family and Len Ainsworth foundation philanthropy. J.J.H. would like to acknowledge funding by the CIHR and CancerCare Manitoba Foundation. P.T. is a National Health and Medical Research Council (NHMRC) Senior Research Fellow. M.N. and D.H. are supported by CINSW Early Career Research Fellowships and St Vincent's Clinic Foundation grants and NHMRC Ideas grants. J.R.W.C. was supported by the European Union's Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement (841973) and an Academy of Finland postdoctoral research grant (338585). O.J.S. and J.P.M. were supported by Cancer Research UK core funding to the Beatson Institute (A31287), to O.J.S. laboratory (A21139), and to J.P.M. laboratory (A29996). B.A.P. and K.J.M. are supported by CINSW Early Career Research Fellowships. B.A.P. is supported by a PanKind (Pancreatic Cancer Foundation) Australia, UNSW Medicine Cancer Theme, Perpetual Impact Philanthropy, and St Vincent's Clinic Foundation grants. Y.-C.S. is supported by an NHMRC grant and the Diabetes Australia Research Program. T.P. is supported by a senior research fellowship through the National Health and Medical Research Council (NHMRC 1155678).

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10.1126/sciadv.adf9063

sciadv.adf9063Slutlig publicerad version, 2,23 MBLicens: CC BY

https://urn.fi/URN:NBN:fi-fe202402136925

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Conway, J. R. W., Warren, S. C., Lee, Y. K., McCulloch, A. T., Magenau, A., Lee, V., Metcalf, X. L., Stoehr, J., Haigh, K., Abdulkhalek, L., Guaman, C. S., Reed, D. A., Murphy, K. J., Pereira, B. A., Mélénec, P., Chambers, C., Latham, S. L., Lenthall, H., Deenick, E. K., ... Timpson, P. (2023). Monitoring AKT activity and targeting in live tissue and disease contexts using a real-time Akt-FRET biosensor mouse. Science Advances, 9(17), Artikel eadf9063. https://doi.org/10.1126/sciadv.adf9063

@article{020e6061dc9b43bca011a781aeabc0bd,

title = "Monitoring AKT activity and targeting in live tissue and disease contexts using a real-time Akt-FRET biosensor mouse",

abstract = "Aberrant AKT activation occurs in a number of cancers, metabolic syndrome, and immune disorders, making it an important target for the treatment of many diseases. To monitor spatial and temporal AKT activity in a live setting, we generated an Akt-FRET biosensor mouse that allows longitudinal assessment of AKT activity using intravital imaging in conjunction with image stabilization and optical window technology. We demonstrate the sensitivity of the Akt-FRET biosensor mouse using various cancer models and verify its suitability to monitor response to drug targeting in spheroid and organotypic models. We also show that the dynamics of AKT activation can be monitored in real time in diverse tissues, including in individual islets of the pancreas, in the brown and white adipose tissue, and in the skeletal muscle. Thus, the Akt-FRET biosensor mouse provides an important tool to study AKT dynamics in live tissue contexts and has broad preclinical applications.",

author = "Conway, \{James R.W.\} and Warren, \{Sean C.\} and Lee, \{Young Kyung\} and McCulloch, \{Andrew T.\} and Astrid Magenau and Victoria Lee and Metcalf, \{Xanthe L.\} and Janett Stoehr and Katharina Haigh and Lea Abdulkhalek and Guaman, \{Cristian S.\} and Reed, \{Daniel A.\} and Murphy, \{Kendelle J.\} and Pereira, \{Brooke A.\} and Pauline M{\'e}l{\'e}nec and Cecilia Chambers and Latham, \{Sharissa L.\} and Helen Lenthall and Deenick, \{Elissa K.\} and Yuanqing Ma and Tri Phan and Elgene Lim and Joshua, \{Anthony M.\} and Stacey Walters and Grey, \{Shane T.\} and Shi, \{Yan Chuan\} and Lei Zhang and Herbert Herzog and Croucher, \{David R.\} and Andy Philp and Scheele, \{Colinda L.G.J.\} and David Herrmann and Sansom, \{Owen J.\} and Morton, \{Jennifer P.\} and Antonella Papa and Haigh, \{Jody J.\} and Max Nobis and Paul Timpson",

note = "Publisher Copyright: {\textcopyright} 2023 The Authors.",

year = "2023",

month = apr,

doi = "10.1126/sciadv.adf9063",

language = "English",

volume = "9",

journal = "Science Advances",

issn = "2375-2548",

publisher = "American association for the advancement of science",

number = "17",

}

Conway, JRW, Warren, SC, Lee, YK, McCulloch, AT, Magenau, A, Lee, V, Metcalf, XL, Stoehr, J, Haigh, K, Abdulkhalek, L, Guaman, CS, Reed, DA, Murphy, KJ, Pereira, BA, Mélénec, P, Chambers, C, Latham, SL, Lenthall, H, Deenick, EK, Ma, Y, Phan, T, Lim, E, Joshua, AM, Walters, S, Grey, ST, Shi, YC, Zhang, L, Herzog, H, Croucher, DR, Philp, A, Scheele, CLGJ, Herrmann, D, Sansom, OJ, Morton, JP, Papa, A, Haigh, JJ, Nobis, M & Timpson, P 2023, 'Monitoring AKT activity and targeting in live tissue and disease contexts using a real-time Akt-FRET biosensor mouse', Science Advances, vol. 9, nr. 17, eadf9063. https://doi.org/10.1126/sciadv.adf9063

TY - JOUR

T1 - Monitoring AKT activity and targeting in live tissue and disease contexts using a real-time Akt-FRET biosensor mouse

AU - Conway, James R.W.

AU - Warren, Sean C.

AU - Lee, Young Kyung

AU - McCulloch, Andrew T.

AU - Magenau, Astrid

AU - Lee, Victoria

AU - Metcalf, Xanthe L.

AU - Stoehr, Janett

AU - Haigh, Katharina

AU - Abdulkhalek, Lea

AU - Guaman, Cristian S.

AU - Reed, Daniel A.

AU - Murphy, Kendelle J.

AU - Pereira, Brooke A.

AU - Mélénec, Pauline

AU - Chambers, Cecilia

AU - Latham, Sharissa L.

AU - Lenthall, Helen

AU - Deenick, Elissa K.

AU - Ma, Yuanqing

AU - Phan, Tri

AU - Lim, Elgene

AU - Joshua, Anthony M.

AU - Walters, Stacey

AU - Grey, Shane T.

AU - Shi, Yan Chuan

AU - Zhang, Lei

AU - Herzog, Herbert

AU - Croucher, David R.

AU - Philp, Andy

AU - Scheele, Colinda L.G.J.

AU - Herrmann, David

AU - Sansom, Owen J.

AU - Morton, Jennifer P.

AU - Papa, Antonella

AU - Haigh, Jody J.

AU - Nobis, Max

AU - Timpson, Paul

PY - 2023/4

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N2 - Aberrant AKT activation occurs in a number of cancers, metabolic syndrome, and immune disorders, making it an important target for the treatment of many diseases. To monitor spatial and temporal AKT activity in a live setting, we generated an Akt-FRET biosensor mouse that allows longitudinal assessment of AKT activity using intravital imaging in conjunction with image stabilization and optical window technology. We demonstrate the sensitivity of the Akt-FRET biosensor mouse using various cancer models and verify its suitability to monitor response to drug targeting in spheroid and organotypic models. We also show that the dynamics of AKT activation can be monitored in real time in diverse tissues, including in individual islets of the pancreas, in the brown and white adipose tissue, and in the skeletal muscle. Thus, the Akt-FRET biosensor mouse provides an important tool to study AKT dynamics in live tissue contexts and has broad preclinical applications.

AB - Aberrant AKT activation occurs in a number of cancers, metabolic syndrome, and immune disorders, making it an important target for the treatment of many diseases. To monitor spatial and temporal AKT activity in a live setting, we generated an Akt-FRET biosensor mouse that allows longitudinal assessment of AKT activity using intravital imaging in conjunction with image stabilization and optical window technology. We demonstrate the sensitivity of the Akt-FRET biosensor mouse using various cancer models and verify its suitability to monitor response to drug targeting in spheroid and organotypic models. We also show that the dynamics of AKT activation can be monitored in real time in diverse tissues, including in individual islets of the pancreas, in the brown and white adipose tissue, and in the skeletal muscle. Thus, the Akt-FRET biosensor mouse provides an important tool to study AKT dynamics in live tissue contexts and has broad preclinical applications.

UR - https://www.scopus.com/pages/publications/85158018642

U2 - 10.1126/sciadv.adf9063

DO - 10.1126/sciadv.adf9063

M3 - Article

C2 - 37126544

AN - SCOPUS:85158018642

SN - 2375-2548

VL - 9

JO - Science Advances

JF - Science Advances

IS - 17

M1 - eadf9063

ER -

Monitoring AKT activity and targeting in live tissue and disease contexts using a real-time Akt-FRET biosensor mouse

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